主 办:北 京 中 医 药 大 学
ISSN 1006-2157 CN 11-3574/R

北京中医药大学学报 ›› 2019, Vol. 42 ›› Issue (5): 389-395.doi: 10.3969/j.issn.1006-2157.2019.05.006

• 中药药理 • 上一篇    下一篇

祛风宣肺方对神经生长因子介导的人支气管上皮细胞神经源性炎症的干预作用及机制*

季坤, 史利卿#, 马建岭, 林彬, 李扭扭, 董尚娟, 温绍惠, 王丽云, 王梁敏   

  1. 北京中医药大学东方医院呼吸热病科 北京 100078
  • 收稿日期:2018-11-08 出版日期:2019-05-30 发布日期:2019-06-05
  • 通讯作者: 史利卿, 男, 博士, 主任医师, 教授, 博士生导师, 研究方向:中医药治疗呼吸系统疾病, E-mail:shiliqing3662@sina.com
  • 作者简介:季坤,女,博士,主治医师
  • 基金资助:
    *国家自然科学基金资助项目(No.81373588),北京市自然科学基金资助项目(No.7184224),北京中医药大学自主选题项目(No.2017-JYBZZ-JS102、 No.2018-JYBZZ-JS103)

Mechanism of Qufeng Xuanfei Fang in intervention of neurogenic inflammation of human bronchial epithelial cells mediated by nerve growth factor*

Ji Kun, Shi Liqing#, MA Jianling, Lin Bin, Li Niuniu, Dong Shangjuan, Wen Shaohui, Wang Liyun, Wang Liangmin   

  1. Department of Respiratory and Fever Diseases, Dongfang Hospital, Beijing University of Chinese Medicine, Beijing 100078, China
  • Received:2018-11-08 Online:2019-05-30 Published:2019-06-05
  • Contact: Prof. Shi Liqing, Ph. D., Chief Physician, Doctoral supervisor. Department of Respiratory and Fever Diseases, Dongfang Hospital, Beijing University of Chinese Medicine, No.6, Area 1, Fangxing Yuan, Fangzhuang, Fengtai District, Beijing 100078. E-mail: shiliqing 3662@sina.com

摘要: 目的 探讨祛风宣肺方对神经生长因子(NGF)介导的人支气管上皮细胞(HBEC)神经源性炎症的干预作用及机制。方法 人支气管上皮细胞随机分为正常组,模型组,阿斯美组,祛风宣肺方高、中、低剂量组,利用NGF制备体外神经源性炎症细胞模型,造模后给予含药血清干预。采用四甲基偶氮唑蓝(MTT)法检测各组细胞的增殖率,使用流式细胞仪检测各组细胞凋亡率,并用RT-PCR及蛋白质印迹(Western blot)法检测各组细胞Pan-Ras、c-fos、神经激肽1受体(NK-1R) mRNA及蛋白表达情况。结果 与正常组相比,模型组细胞增殖率、凋亡率、Pan-Ras、c-fos、NK-1R mRNA及蛋白的表达均升高,差异有统计学意义(P<0.05)。与模型组相比,阿斯美组、祛风宣肺方高、中、低剂量组细胞增殖率、凋亡率、Pan-Ras、c-fos mRNA及蛋白的表达均降低,差异有统计学意义(P<0.05);祛风宣肺方高、中剂量组均能够明显降低NK-1R mRNA及蛋白的表达,差异具有统计学意义(P<0.05)。结论 祛风宣肺方通过下调Ras-Raf-MAPK信号通路相关基因和蛋白的表达,从而减轻NGF介导的气道神经源性炎症,降低咳嗽敏感性。

关键词: 祛风宣肺方, 神经生长因子, 气道神经源性炎症, 慢性咳嗽

Abstract: Objective To investigate the interventional effect and mechanism of Qufeng Xuanfei Fang (Wind-expelling Lung-dispersing Decoction, QFXF Decoction) to neurogenic inflammation of human bronchial epithelial cells (HBEC) mediated by nerve growth factor (NGF). Methods HBEC were randomly divided into normal group, model group, Asmeton group, and high-dose, mid-dose and low-dose QFXF Decoction groups. The model of in vitro neurogenic inflammation was established by using NGF. The normal group was given normal serum, model group, 100 ng/mL of NGF and normal serum, Asmeton group, 100 ng/mL of NGF and Asmeton medicated serum, and high-dose, mid-dose and low-dose QFXF Decoction groups, QFXF Decoction medicated serum in different doses(100 ng/mL of NGF and high-dose, mid-dose or low-dose QFXF Decoction). The proliferation rate of HBEC was detected by using MTT, apoptosis rate was detected by using flow cytometry, and mRNA and protein expressions of Pan-Ras, c-fos and NK-1R were detected by byusing RT-PCR and Western blotting assay. Results The proliferation rate and apoptosis rate of HBEC, and mRNA and protein expressions of Pan-Ras, c-fos and NK-1R increased all in model group compared with normal group(P<0.05). The proliferation rate and apoptosis rate of HBEC, and mRNA and protein expressions of Pan-Ras and c-fos decreased in Asmeton group, and high-dose, mid-dose and low-dose QFXF Decoction groups compared with model group(P<0.05). The mRNA and protein expressions of NK-1R decreased significantly in high-dose and mid-dose QFXF Decoction groups(P<0.05), while they had no changes in Asmeton group and low-dose QFXF Decoction group (P>0.05). Conclusion QFXF Decoction can relieve HBEC neurogenic inflammation mediated by NGF and reduce cough sensitivity through down-regulating the expressions of genes and protein related to Ras-Raf-MAPK signaling pathway.

Key words: Qufeng Xuanfei Fang(Wind-expelling Lung-dispersing Decoction), nerve growth factor, airway neurogenic inflammation, chronic cough

中图分类号: 

  • R285.5